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In this study, screening and identification differentially displayed genes related to breast blisters (keel cysts) in the Gushi chickensxAnkao F2 resource population. Delta Different Display Reverse Transcription-Polymerase Chain Reaction (DDRT-PCR) in combination with Single Strand Conformation Polymorphism (SSCP) is a very sensitive tool for analyzing gene expression of isolated liver tissue from birds. The special primer pair was designed to identify the differential displayed band and a semi-quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR) was performed simultaneously. We now are able to directly characterize genetic differences in breast blisters and the healthy birds. The new ESTs was submitted to GenBank (Accession number: GW317166, GW317167, GW317168, GW317169, GW317170, GW317171, GW836603). Several of the differentially expressed fragments were messenger RNA. Six fragments showed interesting homologies to known sequences from the databases. One fragment did not show any homologies to known sequences. This lays a foundation for further study on the mechanism of differential gene expression in breast blisters and the healthy chickens. It is the first time to demonstrate in this study that Tumor Susceptibility Gene 101 (TSG101) is related to breast blisters (keel cysts) of poultry. Furthermore, GW317167 is suggested to contribute to the development of breast blisters (keel cysts), thus may be candidates of new targets of tumor susceptibility genes.